Cell Biology

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    Protocols in Current Issue
    Tandem Affinity Purification of SBP-CBP-tagged Type Three Secretion System Effectors
    Authors:  Laura Berneking, Marie Schnapp, Theresa Nauth and Moritz Hentschke, date: 06/20/2019, view: 351, Q&A: 0
    [Abstract] Identification of protein-protein interactions of bacterial effectors and cellular targets during infection is at the core to understand how bacteria manipulate the infected host to overcome the immune response. Potential interacting proteins might be identified by genetic methods, i.e., two hybrid screens and could be verified by ...
    Cell Synchronization by Double Thymidine Block
    Authors:  Guo Chen and Xingming Deng, date: 09/05/2018, view: 10885, Q&A: 1
    [Abstract] Cell synchronization is widely used in studying mechanisms involves in regulation of cell cycle progression. Through synchronization, cells at distinct cell cycle stage could be obtained. Thymidine is a DNA synthesis inhibitor that can arrest cell at G1/S boundary, prior to DNA replication. Here, we present the protocol to synchronize cells at ...
    An in vitro Co-culture System for the Activation of CD40 by Membrane-presented CD40 Ligand versus Soluble Agonist
    [Abstract] One fundamental property of the TNR receptor (TNFR) family relates to how ‘signal quality’ (the extent of receptor ligation or cross-linking) influences the outcome of receptor ligation, for instance the induction of death in tumour cells. It is unequivocal that membrane-presented ligand (delivered to target cells via cell-surface presentation by ...
    3D Culture Protocol for Testing Gene Knockdown Efficiency and Cell Line Derivation
    Authors:  Jan Strnadel, Sang Myung Woo, Sunkyu Choi, Huawei Wang, Marian Grendar and Ken Fujimura, date: 06/05/2018, view: 3457, Q&A: 0
    [Abstract] Traditional 2D cell cultures with cells grown as monolayers on solid surface still represent the standard method in cancer research for drug testing. Cells grown in 2D cultures, however, lack relevant cell-matrix and cell-cell interactions and ignore the true three-dimensional anatomy of solid tumors. Cells cultured in 2D can also undergo ...
    Measurement of Intracellular ROS in Caenorhabditis elegans Using 2’,7’-Dichlorodihydrofluorescein Diacetate
    Authors:  Dong Suk Yoon, Myon-Hee Lee and Dong Seok Cha, date: 03/20/2018, view: 4508, Q&A: 1
    [Abstract] Reactive oxygen species (ROS) are generated during normal metabolic processes under aerobic conditions. Since ROS production initiates harmful radical chain reactions on cellular macromolecules, including lipid peroxidation, DNA mutation, and protein denaturation, it has been implicated in a wide spectrum of diseases such as cancer, ...
    Detection of Intracellular Reduced (Catalytically Active) SHP-1 and Analyses of Catalytically Inactive SHP-1 after Oxidation by Pervanadate or H2O2
    Authors:  Seeyoung Choi and Paul E. Love, date: 01/05/2018, view: 2286, Q&A: 0
    [Abstract] Oxidative inactivation of cysteine-dependent Protein Tyrosine Phosphatases (PTPs) by cellular reactive oxygen species (ROS) plays a critical role in regulating signal transduction in multiple cell types. The phosphatase activity of most PTPs depends upon a ‘signature’ cysteine residue within the catalytic domain that is maintained in the ...
    Lentiviral Knockdown of Transcription Factor STAT1 in Peromyscus leucopus to Assess Its Role in the Restriction of Tick-borne Flaviviruses
    Authors:  Adaeze O. Izuogu and R. Travis Taylor, date: 12/05/2017, view: 4125, Q&A: 0
    [Abstract] Cellular infection with tick-borne flaviviruses (TBFVs) results in activation of the interferon (IFN) signaling pathway and subsequent upregulation of numerous genes termed IFN stimulated genes (ISGs) (Schoggins et al., 2011). Many ISGs function to prevent virus pathogenesis by acting in a broad or specific manner through protein-protein ...
    GFP-Grb2 Translocation Assay Using High-content Imaging to Screen for Modulators of EGFR-signaling
    Authors:  Julia Petschnigg and Robin Ketteler, date: 09/05/2017, view: 3514, Q&A: 0
    [Abstract] High-content screening is a useful tool to understand complex cellular processes and to identify genes, proteins or small molecule compounds that modulate such pathways. High-content assays monitor the function of a protein or pathway by visualizing a change in an image-based readout, such as a change in the localization of a reporter protein. ...
    Ciberial Muscle 9 (CM9) Electrophysiological Recordings in Adult Drosophila melanogaster
    Authors:  Benjamin A. Eaton and Rebekah E. Mahoney, date: 07/20/2017, view: 3231, Q&A: 0
    [Abstract] The complexity surrounding presynaptic recordings in mammals is a significant barrier to the study of presynaptic mechanisms during neurotransmission in the mammalian central nervous system (CNS). Here we describe an adult fly neuromuscular junction (NMJ), the ciberial muscle 9 (CM9) NMJ, which allows for the recording of both evoked (EPSPs) and ...
    Measurement of the Intracellular Calcium Concentration with Fura-2 AM Using a Fluorescence Plate Reader
    Authors:  Magdiel Martínez, Namyr A. Martínez and Walter I. Silva, date: 07/20/2017, view: 13409, Q&A: 0
    [Abstract] Intracellular calcium elevation triggers a wide range of cellular responses. Calcium responses can be affected or modulated by membrane receptors mutations, localization, exposure to agonists/antagonists, among others (Burgos et al., 2007; Martínez et al., 2016). Changes in intracellular calcium concentration can be measured ...



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